FIGURE 6:

Filamin-mediated monolayer stiffening facilitates long-term junction mechanoadaptation. (A) Schematic illustration of the atomic force microscopy experiments to quantify monolayer cortical stiffness. (B) Frequency distribution of monolayer elastic moduli show stiffening in response to stretch (n = 150–200 force curves pooled across three independent experiments; ***p < 0.0001, 30 min, 20% versus 0%, **p < 0.005, 3 h, 20% versus 0%, Kolmogorov–Smirnov test). (C) Schematic illustration of the atomic force microscopy experiments to quantify monolayer cortical stiffness in filamin-depleted cells. (D) Frequency distribution of monolayer elastic moduli shows that depletion of filamin A and to a lesser extent filamin B prevents monolayer stiffening in response to stretch (n = 200–250 force curves pooled across three independent experiments; ***p ≤ 0.0001, Kolmogorov–Smirnov test). (E, F) Representative immunofluorescence images, E, and quantification, F, of AJ remodeling from β-catenin show that depletion of filamin A or B prevents full restoration of adherens junction architecture at 3 h of stretch (mean ± SD; n = 3 independent experiments; ***p = 0009, **p = 0.0053, ANOVA with Dunnett’s; scale bars 30 μm). (G) Model of temporal dynamics of AJ remodeling and reinforcement in response to stretch.